title: Neural stem cell isolation from the whole mouse brain using the novel FABP7-binding fluorescent dye, CDr3.
abstract: Methods for the isolation of live neural stem cells from the brain are limited due to the lack of well-defined cell surface markers and tools to detect intracellular markers. To date most methods depend on the labeling of extracellular markers using antibodies, with intracellular markers remaining inaccessible in live cells. Using a novel intracellular protein FABP7 (Fatty Acid Binding Protein-7) selective fluorescent chemical probe CDr3, we have successfully isolated high FABP7 expressing cells from the embryonic and adult mouse brains. These cells are capable of forming neurospheres in culture, express neural stem cell marker genes and differentiate into neurons, astrocytes and oligodendrocytes. Characterization of cells sorted with Aldefluor or antibodies against CD133 or SSEA-1 showed that the cells isolated by CDr3 exhibit a phenotype distinct from the cells sorted with conventional methods. FABP7 labeling with CDr3 represents a novel method for rapid isolation of neural stem cells based on the expression of a single intracellular marker.
authors.affiliation: NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Republic of Singapore; Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium, Agency for Science, Technology and Research, Republic of Singapore.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Leong
authors.forename: Duanting
authors.initials: D
authors.lastname: Zhai
authors.forename: Beomsue
authors.initials: B
authors.lastname: Kim
authors.forename: Seong-Wook
authors.initials: SW
authors.lastname: Yun
authors.forename: Young-Tae
authors.initials: YT
authors.lastname: Chang
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D001896:Boron Compounds / Q000737:chemistry / Q000378:metabolism*; D001921:Brain / Q000166:cytology*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism; D016256:Lewis X Antigen / Q000276:immunology / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D010641:Phenotype; D011485:Protein Binding
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D001896:Boron Compounds; C000588733:CDr3 compound; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005456:Fluorescent Dyes; D006023:Glycoproteins; D016256:Lewis X Antigen; D009419:Nerve Tissue Proteins; D010455:Peptides; C000606024:Prom1 protein, mouse